隐色素
每2
生物
生物钟
时钟
昼夜节律
遗传学
细胞生物学
视交叉上核
野生型
基因
作者
Gizem Cagla Parlak,Bilge Bahar Camur,Seref Gul,Onur Ozcan,Ibrahim Baris,Ibrahim Halil Kavakli
标识
DOI:10.1016/j.jbc.2022.102334
摘要
Abstract
Human clock-gene variations contribute to the phenotypic differences observed in various behavioral and physiological processes such as diurnal preference, sleep, metabolism, mood regulation, addiction, and fertility. However, little is known about the possible effects of identified variations at the molecular level. In this study, we performed a functional characterization at the cellular level of rare CRYPTOCHROME 2 (CRY2) missense variations that were identified from the Ensembl database. Our structural studies revealed that three variations (p.Pro123Leu, p.Asp406His, p.Ser410Ile) are located at the rim of the secondary pocket of CRY2. We show these variants were unable to repress CLOCK/BMAL1-driven transcription in a cell-based reporter assay and had reduced affinity to CLOCK/BMAL1. Furthermore, our biochemical studies indicated that the variants were less stable than the wild-type CRY2, which could be rescued in the presence of Period 2 (PER2), another core clock protein. Finally, we found these variants were unable to properly localize to the nucleus, and thereby were unable to rescue the circadian rhythm in a Cry1−/−Cry2−/− double-knockout mouse embryonic fibroblast cell line. Collectively, our data suggest that the rim of the secondary pocket of CRY2 plays a significant role in its nuclear localization independently of PER2 and in the intact circadian rhythm at the cellular level.
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